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. 2006 Jul;26(14):5544–5557. doi: 10.1128/MCB.02270-05

FIG. 6.

FIG. 6.

The C1 region mediates coactivation via p300. (A) Zac or ZacΔC1 were transfected into LLC-PK1 cells, and different amounts of cell extracts (left lanes, 50 μg; middle lanes, 20 μg; and right lanes, 5 μg) were immunoblotted with α-Zac. Indicated amounts of DNA refer to Zac with ZacΔC1 adjusted appropriately. (B) The indicated doses of Zac or adjusted amounts of ZacΔC1 were cotransfected with the DR or PAL reporter plasmids (2 μg each) into LLC-PK1 cells. (C) Zac or ZacΔC1 was cotransfected with the indicated reporters in the absence or presence of a given amount of p300 (0.5 μg of pCMV-p300-HA) into PA-TU cells. (D) Colony formation assay. Zac (0.2 μg of pRK7Flag-Zac) or ZacΔC1 (0.04 μg of pRK7Flag-ZacΔC1) were cotransfected with a puromycin resistance vector (pRK7Pur) at a ratio of 3:1 into LLC-PK1 and PA-TU cells. Selection (puromycin, 2 μg/ml) began on day 2, and medium was renewed every third day. Colonies were stained with MTT (methylthiazolyldiphenyl-tetrazolium bromide; 1 mg/ml) on day 10 and counted. Growth inhibition by Zac was set to 100%.